JP2019505567A5 - - Google Patents

Claims (19)

A system comprising a first nucleic acid construct and a second nucleic acid construct ,
In here, the first construct comprises a first synthetic coding sequence encoding a first polypeptide sequence comprising the HPV E6 amino acid sequence conjugated to human papillomavirus (HPV) E7 amino acid sequence, Here, the HPV E6 and E7 amino acid sequences are both identified from the wild-type HPV E6 and E7 coding sequences, respectively, by replacing selected codons in the wild-type HPV E6 or E7 coding sequence with synonymous codons, respectively. And the individual synonymous codons have the preference to generate a greater immune response than the corresponding selected codon, wherein the codon substitution is one selected from Table 1 where the first synthetic least be 70% of the codons of a coding sequence, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97 , 98%, 99%, or 100% are synonymous codons in Table 1, and wherein the first synthetic coding sequence is operably linked to a regulatory nucleic acid sequence; and The construct comprises a second synthetic coding sequence encoding a first polypeptide sequence comprising an HPV E6 amino acid sequence conjugated to the HPV E7 amino acid sequence, wherein the HPV E6 and E7 amino acid sequences are wild-type. Substitution of a selected codon in a type HPV E6 or E7 coding sequence with a synonymous codon will both distinguish them from the wild-type HPV E6 and E7 coding sequences, respectively, where the individual synonymous codons are the corresponding selected codons. Have the preference to generate a greater immune response than the codon substitutions, wherein the codon substitutions are selected from Table 1 and wherein the first synthetic code Least be 70% of the codons of the sequence, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, Or 100% are synonymous codons in Table 1, and wherein the first synthetic coding sequence is operably linked to a regulatory nucleic acid sequence; and to a nucleic acid sequence encoding a ubiquitin molecule; And wherein Table 1 is a construct system, wherein:

The construct system according to claim 1, wherein the nucleic acid encoding the ubiquitin molecule comprises at least a part of the sequence shown in SEQ ID NO: 5. 3. The construct system of claim 2, wherein said first synthetic coding sequence comprises the polynucleotide sequence set forth in SEQ ID NO: 7. The construct system according to any one of claims 1 to 3, wherein the second synthetic coding sequence comprises the polynucleotide sequence shown in SEQ ID NO: 8. 5. The method of claim 1, wherein the first synthetic coding sequence comprises the polynucleotide sequence set forth in SEQ ID NO: 7, and the second synthetic coding sequence comprises the polynucleotide sequence set forth in SEQ ID NO: 8. A construct system according to claim 1. The construct system according to any one of claims 1 to 5, wherein the first construct and the second construct are included in one or more expression vectors. The construct system according to any one of claims 1 to 6, wherein the construct system is formulated with a pharmaceutically acceptable carrier or excipient. The construct system according to any one of claims 1 to 7, wherein the construct system is formulated with an adjuvant. For the treatment of human papillomavirus (HPV) infection in a subject, a pharmaceutical composition comprising a construct system comprising a first construct and the second construct, the effective amount of the construct system the subject Administered to
Wherein the first construct comprises a first synthetic coding sequence encoding a first polypeptide sequence comprising an HPV E6 amino acid sequence conjugated to the HPV E7 amino acid sequence, wherein the HPV E6 and The E7 amino acid sequence is both identified from the wild-type HPV E6 and E7 coding sequences, respectively, by replacing selected codons in the wild-type HPV E6 or E7 coding sequence with synonymous codons, where the individual synonymous codons are Has the preference to generate a greater immune response than the corresponding selected codon, wherein the codon substitution is selected from Table 1 and wherein the codon of the first synthetic coding sequence is least be 70% of, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, and 100% are synonymous codons in Table 1 and wherein the first synthetic coding sequence is operably linked to a regulatory nucleic acid sequence; and wherein the second construct is an HPV E7 amino acid sequence A second synthetic coding sequence encoding a first polypeptide sequence comprising the HPV E6 amino acid sequence conjugated to the HPV E6 and E7 amino acid sequences, wherein the HPV E6 and E7 amino acid sequences are in the wild type HPV E6 or E7 coding sequence Replacement of selected codons with synonymous codons, respectively, both discriminate from wild-type HPV E6 and E7 coding sequences, wherein individual synonymous codons are greater in immune response than the corresponding selected codons Wherein the codon substitution is one selected from Table 1 and wherein the codon substitution of the first synthetic coding sequence is Even 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, Table A method of treatment wherein one synonymous codon and wherein the first synthetic coding sequence is operably linked to a regulatory nucleic acid sequence; and to a nucleic acid sequence encoding a ubiquitin molecule. Before administering the first and second constructs simultaneously, the subject is further comprises identifying that it has a H PV infection claim 9 pharmaceutical composition. The pharmaceutical composition according to claim 9 or 10 , wherein the nucleic acid encoding the ubiquitin molecule comprises at least a part of the sequence shown in SEQ ID NO: 5. The pharmaceutical composition according to any one of claims 9 to 11 , wherein the first synthetic coding sequence comprises the polynucleotide sequence shown in SEQ ID NO: 7. The pharmaceutical composition according to any one of claims 9 to 12 , wherein the second synthetic coding sequence comprises the polynucleotide sequence shown in SEQ ID NO: 8. It said first synthetic coding sequence comprises a polynucleotide sequence shown in SEQ ID NO: 7, and a second synthetic coding sequence comprises a polynucleotide sequence shown in SEQ ID NO: 8, claim 9-13 A pharmaceutical composition according to claim 1 . The pharmaceutical composition according to any one of claims 9 to 14 , wherein the first construct and the second construct are contained in one or more expression vectors. The method according to any one of claims 9 to 15 , wherein the first nucleic acid construct comprises the sequence shown in SEQ ID NO: 9, and the second nucleic acid construct comprises the sequence shown in SEQ ID NO: 10. Pharmaceutical composition . 17. The pharmaceutical composition according to any one of claims 9 to 16 , wherein the construct system further comprises an adjuvant. The pharmaceutical composition according to any one of claims 9 to 17 , wherein the subject is a human. Use of the construct system according to any one of claims 1 to 8 for the manufacture of a medicament for the treatment of HPV infection.